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traf3ip2 antibody  (Bio-Techne corporation)


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    Bio-Techne corporation traf3ip2 antibody
    Traf3ip2 Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 90/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    traf3ip2 antibody  (Bio-Techne corporation)
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    Bio-Techne corporation traf3ip2 antibody
    Traf3ip2 Antibody, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    traf3ip2  (Novus Biologicals)
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    Novus Biologicals traf3ip2
    Figure 1. Interleukin-18 (IL-18) upregulates <t>TRAF3IP2</t> but suppresses RECK expression in primary human aortic smooth muscle cells (ASMC). (A–C) IL-18 upregulates TRAF3IP2 expression. ASMCs were grown in complete media, and at 70–80% confluency, made quiescent for 48 h, and then incubated with
    Traf3ip2, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    traf3ip2  (R&D Systems)
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    R&D Systems traf3ip2
    Generation of transgenic mice with cardiomyocyte-specific overexpression of <t>TRAF3IP2</t> (TRAF3IP2-Tg). A, the vector map of α-MyHC-mTRAF3IP2. F and R indicate the binding region of forward and reverse PCR primers that detect an overlapping region of 600 bp. B, electrophoresis of a purified Not1-digested fragment of α-MyHC-mTRAF3IP2-human growth hormone (hGH) poly(A) (7.8 kbp). C, Southern blot analysis of wild type (WT) and TRAF3IP2-Tg mice. Genomic DNA of WT and Tg mice were digested with EcoRV and KpnI and probed with TRAF3IP2 ORF. The predicted band sizes of TRAF3IP2 WT allele are 13.8 and 14.3 kbp after EcoRV and KpnI digestion, respectively (black arrowheads). Tg mice showed two bands after digestion (red arrowheads), indicating that two copies of the transgene were inserted into the Tg mouse genome. D, genotyping was carried out using genomic DNA isolated from tail snips from male and female Tg and control NTg littermates. * denotes transgene with an amplicon size of ∼600 bp.
    Traf3ip2, supplied by R&D Systems, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Figure 1. Interleukin-18 (IL-18) upregulates TRAF3IP2 but suppresses RECK expression in primary human aortic smooth muscle cells (ASMC). (A–C) IL-18 upregulates TRAF3IP2 expression. ASMCs were grown in complete media, and at 70–80% confluency, made quiescent for 48 h, and then incubated with

    Journal: Cells

    Article Title: EF24, a Curcumin Analog, Reverses Interleukin-18-Induced miR-30a or miR-342-Dependent TRAF3IP2 Expression, RECK Suppression, and the Proinflammatory Phenotype of Human Aortic Smooth Muscle Cells.

    doi: 10.3390/cells13201673

    Figure Lengend Snippet: Figure 1. Interleukin-18 (IL-18) upregulates TRAF3IP2 but suppresses RECK expression in primary human aortic smooth muscle cells (ASMC). (A–C) IL-18 upregulates TRAF3IP2 expression. ASMCs were grown in complete media, and at 70–80% confluency, made quiescent for 48 h, and then incubated with

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:400; NB100–56740, Novus Biologicals, LLC, Centennial, CO, USA), Tubulin (1:1000; #2144, Cell Signaling Technology/CST, Danvers, MA, USA), RECK (1 : 1000; #3433, CST), MMP2 (1 : 500; #ab97779, Abcam, Waltham, MA, USA), MMP9 (1 : 1000; #2270, CST), and GAPDH (1:1000; #NB300-221, Novus Biologicals, LLC).

    Techniques: Expressing, Incubation

    Figure 2. Targeting TRAF3IP2 or RECK overexpression blunts IL-18-induced ASMC proliferation and migration. (A–E) Silencing TRAF3IP2 inhibits IL-18-induced ASMC proliferation (B) and migration (C). ASMC were transduced with adenovirus-expressing shRNA targeting human TRAF3IP2 (moi 10 for 48 h), made quiescent, and then exposed to IL-18 (10 ng/mL; experimental design in (A)). ASMC proliferation was assessed after 48 h of IL-18 addition using the CyQUANT Cell proliferation assay (B), and migration after 18 h using Boyden chamber assay (C). ASMCs migrating to the lower surface of the membrane were counted in 10 different fields and summarized as mean ± SEM. (B,C) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+GFP (n = 6). Knockdown of TRAF3IP2 was confirmed by RT-qPCR using a TaqMan™probe (D) and Western blotting (E). (D,E) * p < 0.01 vs. untreated (n = 3). (F,G) Dose-dependent effects of Ad.RECK on RECK expression (experimental design in (F)). Induction of RECK following adenoviral transduction was confirmed by Western blotting with tubulin serving as an internal control (G). (H–K) Forced expression of RECK inhibits IL-18-stimulated ASMC proliferation and migration. ASMCs were transduced with adenovirus-expressing human RECK cDNA (moi 10 for 24 h), made quiescent, and then treated with IL-18 (experimental design in (H)) and analyzed for proliferation (I) and migration (J) as in (B,C). (C,J) The insets show representative images of Matrigel™Transwell invasion. Scale bar: 20 µM. (E,G,K) While a representative immunoblot is shown, the intensities of immunoreactive bands from three (E), four (G) and three (K) independent experiments were semiquantified by densitometry and are summarized on the right. (I) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+GFP (n = 6); (J) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+eGFP (n = 4); (K) * p < 0.01 vs. Untreated; † p < 0.05 vs. IL-18 or IL-18+eGFP (n = 3).

    Journal: Cells

    Article Title: EF24, a Curcumin Analog, Reverses Interleukin-18-Induced miR-30a or miR-342-Dependent TRAF3IP2 Expression, RECK Suppression, and the Proinflammatory Phenotype of Human Aortic Smooth Muscle Cells.

    doi: 10.3390/cells13201673

    Figure Lengend Snippet: Figure 2. Targeting TRAF3IP2 or RECK overexpression blunts IL-18-induced ASMC proliferation and migration. (A–E) Silencing TRAF3IP2 inhibits IL-18-induced ASMC proliferation (B) and migration (C). ASMC were transduced with adenovirus-expressing shRNA targeting human TRAF3IP2 (moi 10 for 48 h), made quiescent, and then exposed to IL-18 (10 ng/mL; experimental design in (A)). ASMC proliferation was assessed after 48 h of IL-18 addition using the CyQUANT Cell proliferation assay (B), and migration after 18 h using Boyden chamber assay (C). ASMCs migrating to the lower surface of the membrane were counted in 10 different fields and summarized as mean ± SEM. (B,C) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+GFP (n = 6). Knockdown of TRAF3IP2 was confirmed by RT-qPCR using a TaqMan™probe (D) and Western blotting (E). (D,E) * p < 0.01 vs. untreated (n = 3). (F,G) Dose-dependent effects of Ad.RECK on RECK expression (experimental design in (F)). Induction of RECK following adenoviral transduction was confirmed by Western blotting with tubulin serving as an internal control (G). (H–K) Forced expression of RECK inhibits IL-18-stimulated ASMC proliferation and migration. ASMCs were transduced with adenovirus-expressing human RECK cDNA (moi 10 for 24 h), made quiescent, and then treated with IL-18 (experimental design in (H)) and analyzed for proliferation (I) and migration (J) as in (B,C). (C,J) The insets show representative images of Matrigel™Transwell invasion. Scale bar: 20 µM. (E,G,K) While a representative immunoblot is shown, the intensities of immunoreactive bands from three (E), four (G) and three (K) independent experiments were semiquantified by densitometry and are summarized on the right. (I) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+GFP (n = 6); (J) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+eGFP (n = 4); (K) * p < 0.01 vs. Untreated; † p < 0.05 vs. IL-18 or IL-18+eGFP (n = 3).

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:400; NB100–56740, Novus Biologicals, LLC, Centennial, CO, USA), Tubulin (1:1000; #2144, Cell Signaling Technology/CST, Danvers, MA, USA), RECK (1 : 1000; #3433, CST), MMP2 (1 : 500; #ab97779, Abcam, Waltham, MA, USA), MMP9 (1 : 1000; #2270, CST), and GAPDH (1:1000; #NB300-221, Novus Biologicals, LLC).

    Techniques: Over Expression, Migration, Transduction, Expressing, shRNA, CyQUANT Assay, Proliferation Assay, Boyden Chamber Assay, Membrane, Knockdown, Quantitative RT-PCR, Western Blot, Control

    Figure 3. TRAF3IP2 knockdown restores SMC marker expression and inhibits ASMC proinflamma- tory phenotype without affecting cell viability. (A–G) Silencing TRAF3IP2 restores IL-18-mediated suppression in SMC markers, but inhibits the expression of proinflammatory phenotype markers, without significantly modulating cell viability. ASMCs were transduced with adenoviral TRAF3IP2 shRNA (moi10 for 48 h), made quiescent and then treated with IL-18 (10 ng/mL for 48 h; experimental design in (A)). Expressions of the SMC markers ACTA2 (B,C) and MYH11 (D,E) were analyzed by both RT-qPCR (B,D) and Western blotting (C,E). The proinflammatory phenotype markers Galectin 3, Olr1, VCAM, CCL2, IL-6, IL-8, and TNF-α were analyzed by RT-qPCR using TaqMan™probes (F). Cell viability was assessed by analyzing cleaved caspase-3 levels using a commercially available Caspase-3 (Cleaved) Human ELISA (G). H2O2 (100 µM for 18 h) served as a positive control and induced a significant increase in cleaved capase-3 levels. (C,E) While a representative immunoblot is shown, the intensities of immunoreactive bands from three independent experiments were semiquan- tified by densitometry and are summarized on the right. (B,D,F,G) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+GFP (n = 6 or 7). (C,E) * p < 0.05 vs. Untreated; † p < 0.05 vs. IL-18 or IL-18+GFP (n = 3).

    Journal: Cells

    Article Title: EF24, a Curcumin Analog, Reverses Interleukin-18-Induced miR-30a or miR-342-Dependent TRAF3IP2 Expression, RECK Suppression, and the Proinflammatory Phenotype of Human Aortic Smooth Muscle Cells.

    doi: 10.3390/cells13201673

    Figure Lengend Snippet: Figure 3. TRAF3IP2 knockdown restores SMC marker expression and inhibits ASMC proinflamma- tory phenotype without affecting cell viability. (A–G) Silencing TRAF3IP2 restores IL-18-mediated suppression in SMC markers, but inhibits the expression of proinflammatory phenotype markers, without significantly modulating cell viability. ASMCs were transduced with adenoviral TRAF3IP2 shRNA (moi10 for 48 h), made quiescent and then treated with IL-18 (10 ng/mL for 48 h; experimental design in (A)). Expressions of the SMC markers ACTA2 (B,C) and MYH11 (D,E) were analyzed by both RT-qPCR (B,D) and Western blotting (C,E). The proinflammatory phenotype markers Galectin 3, Olr1, VCAM, CCL2, IL-6, IL-8, and TNF-α were analyzed by RT-qPCR using TaqMan™probes (F). Cell viability was assessed by analyzing cleaved caspase-3 levels using a commercially available Caspase-3 (Cleaved) Human ELISA (G). H2O2 (100 µM for 18 h) served as a positive control and induced a significant increase in cleaved capase-3 levels. (C,E) While a representative immunoblot is shown, the intensities of immunoreactive bands from three independent experiments were semiquan- tified by densitometry and are summarized on the right. (B,D,F,G) * p < at least 0.01 vs. Untreated; † p < 0.01 vs. IL-18 or IL-18+GFP (n = 6 or 7). (C,E) * p < 0.05 vs. Untreated; † p < 0.05 vs. IL-18 or IL-18+GFP (n = 3).

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:400; NB100–56740, Novus Biologicals, LLC, Centennial, CO, USA), Tubulin (1:1000; #2144, Cell Signaling Technology/CST, Danvers, MA, USA), RECK (1 : 1000; #3433, CST), MMP2 (1 : 500; #ab97779, Abcam, Waltham, MA, USA), MMP9 (1 : 1000; #2270, CST), and GAPDH (1:1000; #NB300-221, Novus Biologicals, LLC).

    Techniques: Knockdown, Marker, Expressing, Transduction, shRNA, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Positive Control

    Figure 7. EF24 reverses IL-18-induced upregulation in TRAF3IP2 expression and RECK suppression. (A–C) EF24 blunts IL-18-induced TRAF3IP2 expression. Quiescent ASMCs were treated with EF24

    Journal: Cells

    Article Title: EF24, a Curcumin Analog, Reverses Interleukin-18-Induced miR-30a or miR-342-Dependent TRAF3IP2 Expression, RECK Suppression, and the Proinflammatory Phenotype of Human Aortic Smooth Muscle Cells.

    doi: 10.3390/cells13201673

    Figure Lengend Snippet: Figure 7. EF24 reverses IL-18-induced upregulation in TRAF3IP2 expression and RECK suppression. (A–C) EF24 blunts IL-18-induced TRAF3IP2 expression. Quiescent ASMCs were treated with EF24

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:400; NB100–56740, Novus Biologicals, LLC, Centennial, CO, USA), Tubulin (1:1000; #2144, Cell Signaling Technology/CST, Danvers, MA, USA), RECK (1 : 1000; #3433, CST), MMP2 (1 : 500; #ab97779, Abcam, Waltham, MA, USA), MMP9 (1 : 1000; #2270, CST), and GAPDH (1:1000; #NB300-221, Novus Biologicals, LLC).

    Techniques: Expressing

    Figure 10. Schematic showing that EF24, a curcumin analog with better bioavailability and biologic activity, inhibits the proinflammatory IL-18-induced primary human aortic smooth muscle cells’ (ASMCs) proliferation, migration, and proinflammatory phenotype changes. EF24 inhibits the stress-activated kinase-dependent miR-30a and miR-342 inhibition, TRAF3IP2 upregulation and RECK suppression. While miR-30a mimic reverses IL-18-induced TRAF3IP2 upregulation, the miR- 342 mimic restores IL-18-mediated RECK suppression, potentially via reduced DNMT1 expression and promoter demethylation (dashed purple box). While IL-18 promotes ASMC migration and proliferation, these effects were reversed by TRAF3IP2 knockdown or the ectopic expression of RECK. Further, while IL-18 induced ASMC migration in part via induction of the gelatinases MMP2 and MMP9, these effects were inhibited by EF24. Moreover, EF24 restores IL-18-mediated suppression in SMC markers and blunts the expression of the proinflammatory phenotype markers. These results suggest that the curcumin analog EF24 reverses IL-18-induced ASMC proliferation and migration and proinflammatory phenotypic changes by targeting TRAF3IP2 and restoring RECK expression. These results suggest the therapeutic potential of EF24 in vascular inflammatory and proliferative diseases, including atherosclerosis.

    Journal: Cells

    Article Title: EF24, a Curcumin Analog, Reverses Interleukin-18-Induced miR-30a or miR-342-Dependent TRAF3IP2 Expression, RECK Suppression, and the Proinflammatory Phenotype of Human Aortic Smooth Muscle Cells.

    doi: 10.3390/cells13201673

    Figure Lengend Snippet: Figure 10. Schematic showing that EF24, a curcumin analog with better bioavailability and biologic activity, inhibits the proinflammatory IL-18-induced primary human aortic smooth muscle cells’ (ASMCs) proliferation, migration, and proinflammatory phenotype changes. EF24 inhibits the stress-activated kinase-dependent miR-30a and miR-342 inhibition, TRAF3IP2 upregulation and RECK suppression. While miR-30a mimic reverses IL-18-induced TRAF3IP2 upregulation, the miR- 342 mimic restores IL-18-mediated RECK suppression, potentially via reduced DNMT1 expression and promoter demethylation (dashed purple box). While IL-18 promotes ASMC migration and proliferation, these effects were reversed by TRAF3IP2 knockdown or the ectopic expression of RECK. Further, while IL-18 induced ASMC migration in part via induction of the gelatinases MMP2 and MMP9, these effects were inhibited by EF24. Moreover, EF24 restores IL-18-mediated suppression in SMC markers and blunts the expression of the proinflammatory phenotype markers. These results suggest that the curcumin analog EF24 reverses IL-18-induced ASMC proliferation and migration and proinflammatory phenotypic changes by targeting TRAF3IP2 and restoring RECK expression. These results suggest the therapeutic potential of EF24 in vascular inflammatory and proliferative diseases, including atherosclerosis.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:400; NB100–56740, Novus Biologicals, LLC, Centennial, CO, USA), Tubulin (1:1000; #2144, Cell Signaling Technology/CST, Danvers, MA, USA), RECK (1 : 1000; #3433, CST), MMP2 (1 : 500; #ab97779, Abcam, Waltham, MA, USA), MMP9 (1 : 1000; #2270, CST), and GAPDH (1:1000; #NB300-221, Novus Biologicals, LLC).

    Techniques: Activity Assay, Migration, Inhibition, Expressing, Knockdown

    Generation of transgenic mice with cardiomyocyte-specific overexpression of TRAF3IP2 (TRAF3IP2-Tg). A, the vector map of α-MyHC-mTRAF3IP2. F and R indicate the binding region of forward and reverse PCR primers that detect an overlapping region of 600 bp. B, electrophoresis of a purified Not1-digested fragment of α-MyHC-mTRAF3IP2-human growth hormone (hGH) poly(A) (7.8 kbp). C, Southern blot analysis of wild type (WT) and TRAF3IP2-Tg mice. Genomic DNA of WT and Tg mice were digested with EcoRV and KpnI and probed with TRAF3IP2 ORF. The predicted band sizes of TRAF3IP2 WT allele are 13.8 and 14.3 kbp after EcoRV and KpnI digestion, respectively (black arrowheads). Tg mice showed two bands after digestion (red arrowheads), indicating that two copies of the transgene were inserted into the Tg mouse genome. D, genotyping was carried out using genomic DNA isolated from tail snips from male and female Tg and control NTg littermates. * denotes transgene with an amplicon size of ∼600 bp.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Generation of transgenic mice with cardiomyocyte-specific overexpression of TRAF3IP2 (TRAF3IP2-Tg). A, the vector map of α-MyHC-mTRAF3IP2. F and R indicate the binding region of forward and reverse PCR primers that detect an overlapping region of 600 bp. B, electrophoresis of a purified Not1-digested fragment of α-MyHC-mTRAF3IP2-human growth hormone (hGH) poly(A) (7.8 kbp). C, Southern blot analysis of wild type (WT) and TRAF3IP2-Tg mice. Genomic DNA of WT and Tg mice were digested with EcoRV and KpnI and probed with TRAF3IP2 ORF. The predicted band sizes of TRAF3IP2 WT allele are 13.8 and 14.3 kbp after EcoRV and KpnI digestion, respectively (black arrowheads). Tg mice showed two bands after digestion (red arrowheads), indicating that two copies of the transgene were inserted into the Tg mouse genome. D, genotyping was carried out using genomic DNA isolated from tail snips from male and female Tg and control NTg littermates. * denotes transgene with an amplicon size of ∼600 bp.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Transgenic Assay, Over Expression, Plasmid Preparation, Binding Assay, Electrophoresis, Purification, Southern Blot, Isolation, Amplification

    Characterization of TRAF3IP2-Tg mice. A and B, TRAF3IP2 mRNA (A) and protein expression (B) in LV tissues of 2-month-old male Tg and NTg mice analyzed by RT-qPCR and immunoblotting. Each lane in panel B represents an individual animal. Densitometric analysis of immunoreactive bands in panel B is summarized on the right. C and D, tissue specificity of transgene-derived TRAF3IP2 overexpression was analyzed by immunoblotting (N, NTg; T, Tg). Densitometric analysis of immunoreactive bands is summarized in D. E, immunofluorescence analysis of TRAF3IP2 overexpression in the heart co-localized with WGA. Scale: 50 μm. TRAF3IP2 overexpression is localized predominantly to cardiomyocytes (CM) but not endothelial or smooth muscle cells of blood vessels (BV). Intensity of TRAF3IP2 immunofluorescence signals were quantified and summarized on the right. F and G, TRAF3IP2 overexpression induces activation of IKKβ, p65, JNK, c-Jun, c/EBPβ, and p38 MAPK in LV tissues from male. Densitometric analysis of immune-reactive bands is summarized on the right (F) but not female Tg (G) as analyzed by immunoblotting using activation-specific antibodies. n = 4/group, Error bars represent S.E. *, p < at least 0.05 versus corresponding NTg.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Characterization of TRAF3IP2-Tg mice. A and B, TRAF3IP2 mRNA (A) and protein expression (B) in LV tissues of 2-month-old male Tg and NTg mice analyzed by RT-qPCR and immunoblotting. Each lane in panel B represents an individual animal. Densitometric analysis of immunoreactive bands in panel B is summarized on the right. C and D, tissue specificity of transgene-derived TRAF3IP2 overexpression was analyzed by immunoblotting (N, NTg; T, Tg). Densitometric analysis of immunoreactive bands is summarized in D. E, immunofluorescence analysis of TRAF3IP2 overexpression in the heart co-localized with WGA. Scale: 50 μm. TRAF3IP2 overexpression is localized predominantly to cardiomyocytes (CM) but not endothelial or smooth muscle cells of blood vessels (BV). Intensity of TRAF3IP2 immunofluorescence signals were quantified and summarized on the right. F and G, TRAF3IP2 overexpression induces activation of IKKβ, p65, JNK, c-Jun, c/EBPβ, and p38 MAPK in LV tissues from male. Densitometric analysis of immune-reactive bands is summarized on the right (F) but not female Tg (G) as analyzed by immunoblotting using activation-specific antibodies. n = 4/group, Error bars represent S.E. *, p < at least 0.05 versus corresponding NTg.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot, Derivative Assay, Over Expression, Immunofluorescence, Activation Assay

    Evaluation of cardiac function in TRAF3IP2-Tg and NTg littermates at 2 months of age by transthoracic echocardiography and cine-MRI. A, a representative M-mode tracing showing anterior wall (AW), internal diameter (ID), and posterior wall (PW). B, B-mode tracings showing epicardium (Ep), endocardium (En), and LV chamber (c). C, structural and functional parameters derived from M- and B-mode tracings (n = 6/group). D, representative mid-ventricle short-axis cine-MRI images corresponding to the end-diastole and end-systole cardiac phases are shown (n = 4–6/group). S, septal wall. Error bars represent S.E. *, p < at least 0.05 versus NTg littermates.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Evaluation of cardiac function in TRAF3IP2-Tg and NTg littermates at 2 months of age by transthoracic echocardiography and cine-MRI. A, a representative M-mode tracing showing anterior wall (AW), internal diameter (ID), and posterior wall (PW). B, B-mode tracings showing epicardium (Ep), endocardium (En), and LV chamber (c). C, structural and functional parameters derived from M- and B-mode tracings (n = 6/group). D, representative mid-ventricle short-axis cine-MRI images corresponding to the end-diastole and end-systole cardiac phases are shown (n = 4–6/group). S, septal wall. Error bars represent S.E. *, p < at least 0.05 versus NTg littermates.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Functional Assay, Derivative Assay

    Anatomic and functional changes in 2-month-old TRAF3IP2-Tg and control NTg littermates n , number of mice; LVID;d, LV internal diameter in diastole; LVID;s, LV internal diameter in systole; LVAW;d, LV anterior wall diameter in diastole; LVPW;d, LV posterior wall diameter at diastole; ESV, end systolic volume; EDV, end diastolic volume; LVEDV, LV end diastolic volume; LVESV, LV end systolic volume; EDP, end diastolic pressure; ESP, end systolic pressure; CO, cardiac output; IFR, initial filling rate; PER, peak ejection rate; PFR, peak filling rate; SV, stroke volume. Results represent the mean ± S.E., and are considered statistically significant if p < 0.05.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Anatomic and functional changes in 2-month-old TRAF3IP2-Tg and control NTg littermates n , number of mice; LVID;d, LV internal diameter in diastole; LVID;s, LV internal diameter in systole; LVAW;d, LV anterior wall diameter in diastole; LVPW;d, LV posterior wall diameter at diastole; ESV, end systolic volume; EDV, end diastolic volume; LVEDV, LV end diastolic volume; LVESV, LV end systolic volume; EDP, end diastolic pressure; ESP, end systolic pressure; CO, cardiac output; IFR, initial filling rate; PER, peak ejection rate; PFR, peak filling rate; SV, stroke volume. Results represent the mean ± S.E., and are considered statistically significant if p < 0.05.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Functional Assay, IF-P

    Cardiac functional changes in 2-month-old female TRAF3IP2-Tg and NTg mice Results represent the mean ± S.E. ( n = 5/group) and are considered statistically significant if p < 0.05. LVID;d, LV internal diameter in diastole; LVID;s, LV internal diameter in systole; LVAW;d, LV anterior wall diameter in diastole; LVPW;d, LV posterior wall diameter at diastole; LVAW;s, LV anterior wall diameter in systole.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Cardiac functional changes in 2-month-old female TRAF3IP2-Tg and NTg mice Results represent the mean ± S.E. ( n = 5/group) and are considered statistically significant if p < 0.05. LVID;d, LV internal diameter in diastole; LVID;s, LV internal diameter in systole; LVAW;d, LV anterior wall diameter in diastole; LVPW;d, LV posterior wall diameter at diastole; LVAW;s, LV anterior wall diameter in systole.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Functional Assay, IF-P

    Transgenic overexpression of TRAF3IP2 results in spontaneous development of myocardial hypertrophy. A, gross morphology of the heart (upper panel) and heart weight (HW) to body weight (BW) ratios (lower panel) in 2-month-old NTg and Tg mice. B, cardiomyocyte cross-sectional area was analyzed using WGA-stained heart sections. Scale: 50 μm. The mean area of cardiomyocytes was quantified and summarized on the right. C, mRNA expression of fetal genes (ANP, BNP, and β-MyHC) in LV tissues as analyzed by RT-qPCR. 18S served as an internal control. D, protein expression of fetal genes and GATA4 was analyzed by immunoblotting. Densitometric analysis of immunoreactive bands is summarized on the right. E, temporal changes in heart weight to body weight ratios. F, ANP and GATA4 expression in female Tg hearts (n = 4/group). Error bars represent S.E. *, p < 0.05; **, p < 0.01; ***, p < 0.001 versus NTg littermates.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Transgenic overexpression of TRAF3IP2 results in spontaneous development of myocardial hypertrophy. A, gross morphology of the heart (upper panel) and heart weight (HW) to body weight (BW) ratios (lower panel) in 2-month-old NTg and Tg mice. B, cardiomyocyte cross-sectional area was analyzed using WGA-stained heart sections. Scale: 50 μm. The mean area of cardiomyocytes was quantified and summarized on the right. C, mRNA expression of fetal genes (ANP, BNP, and β-MyHC) in LV tissues as analyzed by RT-qPCR. 18S served as an internal control. D, protein expression of fetal genes and GATA4 was analyzed by immunoblotting. Densitometric analysis of immunoreactive bands is summarized on the right. E, temporal changes in heart weight to body weight ratios. F, ANP and GATA4 expression in female Tg hearts (n = 4/group). Error bars represent S.E. *, p < 0.05; **, p < 0.01; ***, p < 0.001 versus NTg littermates.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Transgenic Assay, Over Expression, Staining, Expressing, Quantitative RT-PCR, Western Blot

    Transgenic overexpression of TRAF3IP2 results in spontaneous development of cardiac fibrosis. A, mRNA expression of ColIα1, ColIIIα1, and CTGF in LV tissues of 2-month-old Tg and NTg mice as analyzed by RT-qPCR (18S served as an internal control). B, protein levels of ColIα1, ColIIIα1, LOX, and CTGF in LV tissues from Tg and NTg mice as analyzed by immunoblotting. Densitometric analysis of immunoreactive bands is summarized on the right. C, deposition and co-localization of ColIα1 and ColIIIα1 were analyzed in LV tissues from Tg and NTg mice by immunofluorescence. Relative fluorescent signals were quantified and summarized on the right. D, temporal changes in cardiac fibrosis as analyzed by Masson's trichrome staining. Mean collagen positive area is summarized on the right (n = 4/group). Error bars represent S.E. *, p < at least 0.05 versus NTg littermates.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Transgenic overexpression of TRAF3IP2 results in spontaneous development of cardiac fibrosis. A, mRNA expression of ColIα1, ColIIIα1, and CTGF in LV tissues of 2-month-old Tg and NTg mice as analyzed by RT-qPCR (18S served as an internal control). B, protein levels of ColIα1, ColIIIα1, LOX, and CTGF in LV tissues from Tg and NTg mice as analyzed by immunoblotting. Densitometric analysis of immunoreactive bands is summarized on the right. C, deposition and co-localization of ColIα1 and ColIIIα1 were analyzed in LV tissues from Tg and NTg mice by immunofluorescence. Relative fluorescent signals were quantified and summarized on the right. D, temporal changes in cardiac fibrosis as analyzed by Masson's trichrome staining. Mean collagen positive area is summarized on the right (n = 4/group). Error bars represent S.E. *, p < at least 0.05 versus NTg littermates.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Transgenic Assay, Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining

    Transgenic overexpression of TRAF3IP2 results in cardiac fibroblast activation. A, co-localization of CD-31 and α-SMA in hearts of 2-month-old male NTg and Tg mice. Mean fluorescent intensities were quantified and summarized on the right. B, co-localization of tensin and α-SMA. Mean fluorescent intensities were quantified and summarized on the right. C, co-localization of paxillin and α-SMA. Mean fluorescent intensities were quantified and summarized on the right. Blood vessels, n = 4/group. Error bars represent S.E. *, p < 0.05; **, p < 0.005 versus NTg littermates.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Transgenic overexpression of TRAF3IP2 results in cardiac fibroblast activation. A, co-localization of CD-31 and α-SMA in hearts of 2-month-old male NTg and Tg mice. Mean fluorescent intensities were quantified and summarized on the right. B, co-localization of tensin and α-SMA. Mean fluorescent intensities were quantified and summarized on the right. C, co-localization of paxillin and α-SMA. Mean fluorescent intensities were quantified and summarized on the right. Blood vessels, n = 4/group. Error bars represent S.E. *, p < 0.05; **, p < 0.005 versus NTg littermates.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Transgenic Assay, Over Expression, Activation Assay

    Protein Quantibody Array data showing changes in protein levels of various proinflammatory mediators in LV homogenates from 2-month-old male and female TRAF3IP2-Tg mice Results are expressed as -fold change in the Tg compared to corresponding NTg littermates. Proteins that showed a 1.5-fold change or more are depicted. ↑ and ↓ indicate up- or down-regulation, respectively. MCSF, macrophage-colony stimulating factor; TWEAK, TNF-related weak inducer of apoptosis; TWEAKR, TWEAK receptor; BLC, B-lymphocyte chemoattractant; MAdCAM-1, mucosal vascular addressin cell adhesion molecule 1; VCAM-1, vascular cell adhesion molecule 1; β-FGF, basic fibroblast growth factor; ACE, angiotensin converting enzyme; Ang, angiotensin; ECM, extracellular matrix.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Protein Quantibody Array data showing changes in protein levels of various proinflammatory mediators in LV homogenates from 2-month-old male and female TRAF3IP2-Tg mice Results are expressed as -fold change in the Tg compared to corresponding NTg littermates. Proteins that showed a 1.5-fold change or more are depicted. ↑ and ↓ indicate up- or down-regulation, respectively. MCSF, macrophage-colony stimulating factor; TWEAK, TNF-related weak inducer of apoptosis; TWEAKR, TWEAK receptor; BLC, B-lymphocyte chemoattractant; MAdCAM-1, mucosal vascular addressin cell adhesion molecule 1; VCAM-1, vascular cell adhesion molecule 1; β-FGF, basic fibroblast growth factor; ACE, angiotensin converting enzyme; Ang, angiotensin; ECM, extracellular matrix.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques:

    Transgenic overexpression of TRAF3IP2 results in spontaneous induction of pro-inflammatory cytokines and macrophage infiltration. A, mRNA and B, protein expression of IL-6 and IL-18 as analyzed by RT-qPCR and immunoblotting, respectively. The immunoreactive bands in B were semiquantified and summarized on the right. C, macrophage infiltration into heart tissues was analyzed by Mac3 staining, and the fluorescence signals are summarized on the right. Scale, 100 μm. The inset shows a 60× magnification. n = 4/group. Error bars represent S.E. *, p < at least 0.05 versus NTg littermates.

    Journal: The Journal of Biological Chemistry

    Article Title: Cardiac-restricted Overexpression of TRAF3 Interacting Protein 2 (TRAF3IP2) Results in Spontaneous Development of Myocardial Hypertrophy, Fibrosis, and Dysfunction *

    doi: 10.1074/jbc.M116.724138

    Figure Lengend Snippet: Transgenic overexpression of TRAF3IP2 results in spontaneous induction of pro-inflammatory cytokines and macrophage infiltration. A, mRNA and B, protein expression of IL-6 and IL-18 as analyzed by RT-qPCR and immunoblotting, respectively. The immunoreactive bands in B were semiquantified and summarized on the right. C, macrophage infiltration into heart tissues was analyzed by Mac3 staining, and the fluorescence signals are summarized on the right. Scale, 100 μm. The inset shows a 60× magnification. n = 4/group. Error bars represent S.E. *, p < at least 0.05 versus NTg littermates.

    Article Snippet: The following antibodies were used: TRAF3IP2 (1:500; #NB100-56740, Novus), ANP (1:400; #NBP2-14873, Novus), IL-6 (1:200; #AF-406-NA, R&D Systems), α-tubulin (1:1000; #2144, Cell Signaling Technology or CST), phospho-p65 (Ser 536 ; 1:1000; #3031, CST), p65 (1:1000; #8242, CST), phospho-c-Jun (Ser 63 , 1:1000; #9261, CST), c-Jun (1:1000; #9165, CST), JNK (1:1000; #9252, CST), phospho-JNK (Thr 183 /Tyr 185 ; 1: 1000, #4668, CST), IKKβ (1:1000; #2678, CST), and phospho-IKKα/β (Ser 176 / 180 ; 1:1000; #2694, CST), LOX (1:500; #sc-373995, Santa Cruz Biotechnology or SCB), IL-18 (1:100; #sc-7954, SCB), BNP (1:400, #sc-67455, SCB), ColIα1 (1:2000; #ab34170, Abcam), ColIIIα1 (1:2000; #ab7778, Abcam), CTGF (1:500; #210303, United States Biological), and β-MyHC (1:400, #M9850-11C, United States Biological).

    Techniques: Transgenic Assay, Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Staining, Fluorescence